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Literature summary for 2.8.3.B1 extracted from

  • Kim, J.; Darley, D.; Selmer, T.; Buckel, W.
    Characterization of (R)-2-hydroxyisocaproate dehydrogenase and a family III coenzyme A transferase involved in reduction of L-leucine to isocaproate by Clostridium difficile (2006), Appl. Environ. Microbiol., 72, 6062-6069.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.8.3.B1

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli as N- and C-terminal Strep tag II fusion protein Clostridioides difficile

Protein Variants

Protein Variants Comment Organism
D171A mutant enzyme is more than 2000fold less active than the wild-type enzyme Clostridioides difficile
D171N mutant enzyme is more than 2000fold less active than the wild-type enzyme Clostridioides difficile

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
 2 * 43000, SDS-PAGE Clostridioides difficile
86000
-
 gel filtration Clostridioides difficile

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4-methylpent-2-enoyl-CoA + (R)-2-hydroxy-4-methylpentanoate Clostridioides difficile the enzyme is involved in the reductive branch of L-leucine fermentation 4-methylpent-2-enoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?
4-methylpent-2-enoyl-CoA + (R)-2-hydroxy-4-methylpentanoate Clostridioides difficile DSM 1296T the enzyme is involved in the reductive branch of L-leucine fermentation 4-methylpent-2-enoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?

Organism

Organism UniProt Comment Textmining
Clostridioides difficile Q5U921
-
-
Clostridioides difficile DSM 1296T Q5U921
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Clostridioides difficile

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylpent-2-enoyl-CoA + (R)-2-hydroxy-4-methylpentanoate the enzyme is involved in the reductive branch of L-leucine fermentation Clostridioides difficile 4-methylpent-2-enoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?
4-methylpent-2-enoyl-CoA + (R)-2-hydroxy-4-methylpentanoate i.e. isocaprenoyl-CoA. Acetyl-CoA or butyryl-CoA, are not accepted as substrates. The enzymatically in situ-produced 2-isocaprenoyl-CoA is accepted as a substrate of the CoA transferase and mediates the tranfer to (R)-2-hydroxy-4-methylpentanoate. CoA transfers from chemically synthesized (E)-2-isocaprenoyl-CoA to (R)-2-hydroxyisocaproate or to isocaproate can not be observed Clostridioides difficile 4-methylpent-2-enoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 r
4-methylpent-2-enoyl-CoA + (R)-2-hydroxy-4-methylpentanoate the enzyme is involved in the reductive branch of L-leucine fermentation Clostridioides difficile DSM 1296T 4-methylpent-2-enoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 ?
4-methylpent-2-enoyl-CoA + (R)-2-hydroxy-4-methylpentanoate i.e. isocaprenoyl-CoA. Acetyl-CoA or butyryl-CoA, are not accepted as substrates. The enzymatically in situ-produced 2-isocaprenoyl-CoA is accepted as a substrate of the CoA transferase and mediates the tranfer to (R)-2-hydroxy-4-methylpentanoate. CoA transfers from chemically synthesized (E)-2-isocaprenoyl-CoA to (R)-2-hydroxyisocaproate or to isocaproate can not be observed Clostridioides difficile DSM 1296T 4-methylpent-2-enoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 r
4-methylpentanoyl-CoA + (R)-2-hydroxy-4-methylpentanoate i.e. isocaproyl-CoA. Acetyl-CoA or butyryl-CoA, are not accepted as substrates. CoA transfers from chemically synthesized (E)-2-isocaprenoyl-CoA to (R)-2-hydroxyisocaproate or to isocaproate can not be observed Clostridioides difficile 4-methylpentanoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 r
4-methylpentanoyl-CoA + (R)-2-hydroxy-4-methylpentanoate i.e. isocaproyl-CoA. Acetyl-CoA or butyryl-CoA, are not accepted as substrates. CoA transfers from chemically synthesized (E)-2-isocaprenoyl-CoA to (R)-2-hydroxyisocaproate or to isocaproate can not be observed Clostridioides difficile DSM 1296T 4-methylpentanoate + (R)-2-hydroxy-4-methylpentanoyl-CoA
-
 r

Subunits

Subunits Comment Organism
homodimer 2 * 43000, SDS-PAGE Clostridioides difficile

Synonyms

Synonyms Comment Organism
HadA
-
 Clostridioides difficile
isocaprenoyl-CoA:2-hydroxyisocaproate CoA-transferase
-
 Clostridioides difficile

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Clostridioides difficile

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Clostridioides difficile

General Information

General Information Comment Organism
metabolism the enzyme is involved in the reductive branch of L-leucine fermentation Clostridioides difficile